骨髓胚胎样干细胞体内重建dystrophin的表达  

Expression of dystrophin reconstructed by bone marrow embryonic-like stem cells in vivo

作　　者：庞荣清[1] 王强[1] 杨建勇[1] 朱光旭[1] 李自安[1] 何洁[1] 朱向情[1] 阮光萍[1] 潘兴华[1] 

机构地区：[1]成都军区昆明总医院细胞生物治疗中心干细胞与免疫细胞生物医药技术国家地方联合工程实验室云南省细胞治疗技术转化医学重点实验室云南省干细胞工程实验室,昆明650032

出　　处：《西南国防医药》2015年第8期823-825,共3页Medical Journal of National Defending Forces in Southwest China

基　　金：云南省基金项目(2011HB050;2013DA004;2013CA005);成都军区基金项目(2012WJ003);国家自然科学基金项目(31172170)

摘　　要：目的观察骨髓胚胎样干细胞体内再生dystrophin的可行性。方法采用SSP-PCR方法鉴定mdx小鼠的基因型。分离扩增人骨髓胚胎样干细胞,采用DIR标记后,注射细胞到mdx小鼠腹股沟三角皮下,采用活体成像法观察植入细胞的存活情况,采用免疫荧光染色法检测dystrophin的表达。结果杂合子鼠交配可以产生3个基因型的子代鼠,采用SSP-PCR可以鉴定出mdx小鼠基因型;采用无血清培养基可从骨髓中分离到表达SSEA-4和FZD-9的胚胎样干细胞,可在注射细胞的mdx小鼠离体后肢肌肉检测到明显的荧光信号,并在肌肉组织中检测到人dystrophin的表达。结论人骨髓胚胎样干细胞可在mdx小鼠体内存活并表达dystrophin。Objective To observe the feasibility of the dystrophin regeneration by bone marrow embryonic-like stem cells （ELSCs） in vivo. Methods SSP-PCR was used to identify the mdx mice＇s genotype. Human bone marrow embryonic-like stem ceils were separated, isolated, and then after the marking by DIR, subcutaneously injected into the mdx mice＇s inguinal triangle. The survival of implanted cells was observed by living-imaging method, and the dystrophin expression was detected by immunofluorescent staining method. Results The copulation of heterozygous mice could breed offspring with three kinds of genotype. The genotype of mdx mice could be identified by SSP-PCR. ELSCs expressing SSEA-4 and FZD-9 could be isolated from the bone marrow through serum-free medium. Significant fluorescence signal was detected in the ex vivo posterior limb muscle of the mdx mice that were injected with the cells, and the expression of dystrophin was detected in the muscle tissues. Conclusion Human bone marrow embryonic-like stem cells can survive within mdx mice and express dystrophin.

关 键 词：骨髓 胚胎样干细胞 mdx小鼠 基因型 DYSTROPHIN DUCHENNE型肌营养不良 

分 类 号：R318.1[医药卫生—生物医学工程]